Cannabis hydrolysis

The aim of this review is to present the different methodologies reported for cannabis detection in human hair (sample, decontamination, hydrolysis, internal standards [IS], extraction, derivatization, system of detection) and the analytical performances observed. 

Like the hydrolysis methods, many extraction procedures have been described. Table 3 presents the various procedures and the compounds used as IS. With the development of the technology and the introduction of gas chromatography coupled with mass spectrometry (GC/MS), deuterated IS have been used for the quantification of cannabis in hair. Presently, this technique represents the state of the art. Most authors use liquid-liquid extraction only Moeller and Moeller et al. - have reported solid-phase extraction (SPE) procedures. 

Cannabis constituents (alkaline hydrolysis of urine followed by SPE )... 

Cannabidiol (CBD) is a non-psychotropic component of cannabis with possible therapeutic use as an anti-inflammatory drug. Recent studies on both enantiomers of CBD showed enantioselectivity in their interaction with cannabinoid and vanniloid (VRl) receptors as well as on the cellular uptake and enzymatic hydrolysis of anandamide (Bisogno et al. 2001). 

Kelly et al. intravenously administered 5 mg of THC to eight males and periodically monitored THC, THCCOOH, and THCCOOH-glucuronide conjugates by GC/MS [limit of detection (LOD) 1 ng/ml for THC and THCCOOH] in plasma with and without alkaline hydrolysis for up to 10 h, and then once daily for up to 12 days (Kelly and Jones 1992). The elimination half-lives of THC, THCCOOH, and THCCOOH-glucuronide in the plasma of frequent cannabis users were 116.8 min, 5.2 days, and 6.8 days, respectively, and 93.3 min, 6.2 days and 3.7 days in infrequent users. Conjugated THCCOOH was detected in the plasma of 75% of the frequent and 25% of the infrequent users at day 12. 

The most widespread abuse of cannabis is by smoking. It occasionally may be abused orally. When smoked, initial metabolism occurs in the lungs, whereas this takes place in the liver when marijuana is taken orally. The identification of 9-carboxy-tetrahydrocannabinol (THC) in urine is considered to be the best indication of previous cannabis consumption. Before SPE, alkaline hydrolysis of the sample is necessary to free the metabolite 9-carboxy-THC. Afterward, M,0-bis-trimethylsilyl-trifluoroacetamide (BSTFA) and trimethylchlorodilsne (TMCS) are added for derivatization the product is analyzed by GC-MS in El or negative-ion chemical ionization (NICI) mode. ... 

Various hydrolases are in fact associated with the protein bodies, though we should realize that some of the enzymes may be contaminants from the cytoplasm. Acid proteinases have been found in protein bodies of Gossypium [47, 79], V faba [54], Hordeum [55] and Cannabis [66]. In the last, the proteinase is active against the crystalloid moiety (edestin) and has, therefore, been called edestinase [66]. The occurrence of these various enzymes for the hydrolysis of the native protein therefore suggests the possible lysosomal nature of protein bodies. 

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